Compositions and Methods of Dust Control

ABSTRACT

Compositions and methods for producing materials for construction and for dust control utilizing enzyme producing cells, an amount of a nitrogen source such as urea, and an amount of calcium such as calcium chloride. Calcium contributes to the formation of calcium carbonate which creates a solid structure, layer or shield. One or more compositions containing components of the invention can be sprayed or otherwise applied to surfaces for erosion control, foundation support, prevention of sink hole formation, prevention of dust formation, or other applications. Ammonia, water and other by-products of the process can be recycled and re-utilized for the same or other purposes including, for example, as fertilizers and energy sources, or independently fermented from selectively cultivated microorganisms.

REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of U.S. application Ser. No.15/066,692 filed Mar. 10, 2016, which claims priority to U.S.Provisional Application No. 62/200,288 filed Aug. 3, 2015, U.S.Provisional Application No. 62/188,556 filed Jul. 3, 2015, and U.S.Provisional Application No. 62/130,854 filed Mar. 10, 2015, the entiretyof each of which is specifically incorporated by reference.

BACKGROUND 1. Field of the Invention

The invention is directed to kits, compositions, tools and methods fordust control. More particularly, the invention is directed to materialsand methods for dust suppression, with isolated enzymes,enzyme-producing bacteria or cells, or spores that give rise toenzyme-producing microorganisms.

2. Description of the Background

The built environment is primarily constructed using a limited paletteof traditional materials: clay, concrete, glass, steel, and wood.Commonly used throughout history, masonry construction continues to makeup a large part of the built environment, utilized for both load bearingstructures and veneer construction. According to Chaisson, globally,traditional clay brick manufacturing produces over 1.23 trillion unitsper annum with a heavy dependency on non-renewable natural resources.Clay brick manufactured in coal-powered kilns emits approximately 1.3pounds of carbon dioxide per unit. According to Burke, in total, brickmanufacturing emits over 800 million tons of man-made CO₂ each year, andyet represents only one material currently used in buildingconstruction.

Fired clay bricks can be manufactured between 3-20 days, depending onthe equipment and processes used. This range represents modern automatedfactories able to process bricks without manual labor, to the clampmethod of bricks stacked around a burning fire used in many developingnations.

As an alternative to load bearing fired clay masonry, Concrete MasonryUnits [CMU] are widely used as they are more economical, faster tomanufacture and can serve as a structural typology for globalconstruction. Comprised of concrete, these units are made with Portlandcement, large aggregate and sand filler. According to Hanley of theUnited States Environmental Protection Agency, global carbon dioxide(CO₂) emissions from cement production were approximately 829 millionmetric tons of CO₂ in 2000.

These traditional materials contain a high-embodied energy, withcomponents of concrete and steel mined from non-renewable resources.Approximately, forty-percent of global carbon dioxide is linked to theconstruction industry, primarily due to material production anddisposal. Biologically grown materials can be pollution free and containa low embodied energy, if produced as part of a local ecosystem.

Natural cement is created through chemical deposition and chemicalprocesses associated with weathering, and can be found in variouslocations on the earth's crust. The formation of natural sandstones isprimarily attributed to the precipitation of calcite cement. As analternatively to natural deposition, a form of natural cement has beenproduced with urease producing Sporosarcina pasteurii, a nonpathogenic,common-soil bacterium has the ability to induce the production ofcalcite through a chemical reaction. The result is a hardened materialformed in a process referred to by Stocks-Fischer as microbial inducedcalcite precipitation [MICP]. Applications include environmentalstabilization of contaminated soils and encapsulation of hazardous andother contaminants in natural soils and acid mine tailings. Ramachandranand Jonkers have proposed the use of microbes to remediate cracks inconcrete structures and the repair of cracks in monuments. According toDeJong and Whiffin, civil engineering researchers in the United States,Australia and the Netherlands have proposed the use of MICP for soilstabilization and erosion control.

A need exists for a process to manufacture building materials that doesnot impose the high energy costs associated with the manufacture of claybricks and other conventional stone replacement, but utilizes readilyavailable materials and is both economical and environmentally safe.

SUMMARY OF THE INVENTION

The present invention overcomes problems and disadvantages associatedwith current strategies and designs, and provides new tools,compositions, and methods for the manufacture of building materials.

One embodiment of the invention is directed to compositions comprising asupport material to which is coupled urease-producing cells or cellspores and a transport medium and optionally a nutrient mix. Preferablythe support material is organic or inorganic and comprises rock, glass(e.g. Poraver), wood, paper, metal, plastic, polymers, minerals orcombinations thereof. Preferably the composition is a liquid, a gel, asludge, a pump-able slurry, a dry powder or crystals and the supportmaterial is in the form of beads, grains, rods, strands, fibers, flakes,pulverized or crushed stone, crystals, fines, or combinations thereof.Preferably the support material is sand, glass, wood (e.g., residuals,pulp, sawdust, lignin), metal, polymers, fines (e.g., microcellulose),waste materials (e.g., ash, scrubber waste, residuals), co-culturedmicroorganisms or combinations thereof and the urease-producing cells orcell spores comprise yeast, algae, bacteria or eukaryotic cells, cellspores, anaerobic cells, or facultative anaerobic cells. Preferredbacteria are Sporosarcina pasteurii, Bacillus megaterium, Sporosarcinaureae, Proteus vulgaris, Bacillus sphaericus, Myxococcus xanthus,Proteus mirabilis, Helicobacter pylori, or variants, serotypes,mutations or combinations thereof, and preferred yeast, algae, bacteriaor eukaryotic cells or cell spores are genetically engineered. Thesupport material and the cells are preferably coupled via hydrophobicbonds, hydropyllic bonds, ionic bonds, non-ionic bonds, covalent bonds,van der Waal forces, or a combination thereof and/or the supportmaterial is at least partially or totally encompassed by a film thatpromotes binding of the urease-producing cells. Preferred films comprisea polymer or a cell nutrient and preferably the composition contains acoloring agent which may be red, blue, green, yellow or any combinationor shade thereof. Preferably the composition contains an identifyingagent or a detectable marker such as a microscopic tag, a color, anucleic acid or peptide, an enzyme or another substance.

Another embodiment of the invention is directed to kits formanufacturing solid forms comprising: the composition of the invention,a second composition containing nutrients for proliferation of theureases-producing cells and/or germination of the cell spores; aplurality of sets of formworks wherein each set encloses the shape of atleast one solid form and contains one or more porous panels; and a thirdcomposition comprising a calcium source (e.g., CaCl₂), a nitrogen source(e.g., urea) or both a calcium source and a nitrogen source. Preferablythe kit is for the creation of solid forms such as, for example,rectangular, square, rounded, oval or an irregular shape. Preferredsolid forms include but are not limited to blocks, boards, bricks,pavers, panels, tiles, or veneer. Preferably kits of the invention arefor the manufacture of blocks such as, for example, concrete masonry,cinder blocks, foundation blocks, breeze blocks, hollow blocks, solidblocks, besser blocks, clinker blocks, high or low density blocks, oraerated blocks. Preferably the nutrients include amino acids, proteins,polysaccharides, fatty acids, vitamins and minerals.

Another embodiment of the invention is directed to methods formanufacture of solid forms comprising: mixing the composition of theinvention with an aggregate material and water to form a mixture,wherein the aggregate material is largely composed of particulates of 5mm or greater or particles of than 5 mm or less in diameter (e.g.,fines); optionally apportioning the mixture into multiple form workswherein each form work contains at least one porous panel; adding asecond composition to the mixture, wherein the second compositioncontains nutrients that promote proliferation of the urease-producingcells; adding a third composition to the mixture, wherein the thirdcomposition is a liquid that contains calcium; incubating the mixturefor a period of time to form covalent bonds between the particulates;and removing the solid forms from the form works. Preferably theaggregate material comprises rock, glass, wood, paper, metal, plastic,polymers, minerals or combinations thereof, and/or mixing comprisingspraying the composition as a liquid onto the aggregate material.Preferably the form works are substantially submerged during theincubating and air is bubbled to the submerged form works. Preferably athird composition is added to the mixture repeatedly during incubatingwhich drains through the bottom panel and, optionally, is recycled.Preferably, incubating is performed under ambient conditions and thethird composition contains calcium chloride, calcium acetate, calciumphosphate, calcium carbonate, calcium lactate, calcium nitrate, or acalcium salt. Preferably the pH of the mixture is monitored during theincubating. Preferably the solid forms are blocks, boards, bricks, thinbricks, pavers, panels, tiles, or veneer, stone (manufactured, cultured,colored), and the mixture further contains fibers or nanofibers thatare, for example, fibers or nanofibers of wood, glass, plastic, metal ora polymer. Preferred fibers include, for example, polypropylene, HDPE,carbon fibers including high-strength carbon fibers, rayon, andbiodegradable and non-biodegradable fibers such polymers of, forexample, poly lactic acid, fibers of cellulose, minerals, chitin,lignin, and other plant materials. Preferably additional nutrients areadded during incubating and the solid forms removed from the form worksare dried.

Another embodiment of the invention comprises compositions containingurease producing cells or cell spores that are encapsulated or coatedwith nutrient media such as, for example, proteins or polysaccharides,or polymers such as poly lactic acid which is water soluble. Preferablythe nutrient media further contains additional urease producing cells orcell spores.

Another embodiment of the invention comprises methods of dust controlcomprising: providing two compositions, wherein one composition containsan aqueous or dry mixture of viable cells or spores of urease-producingmicroorganisms and a transport media, and the other composition containsan aqueous form of calcium, wherein either or both compositions furthercontain urea and ingredients for growth of the microorganisms; andseparately applying the two compositions to an area. Preferably themicroorganisms comprise yeast, algae, bacteria, eukaryotic cells, orrecombinantly engineered microorganisms, and also preferably thebacteria comprise Sporosarcina pasteurii, Bacillus megaterium,Sporosarcina ureae, Proteus vulgaris, Bacillus sphaericus, Myxococcusxanthus, Proteus mirabilis, Helicobacter pylori, or variants, serotypes,mutations, or combinations thereof. Preferably the transport mediummaintains viability without promoting significant propagation of themicroorganisms and may comprises minimal cell medium. Preferably thecalcium comprises calcium chloride, calcium acetate, calcium phosphate,calcium carbonate, calcium lactate, calcium nitrate, and/or a calciumsalt, and the ingredients for growth comprise one or more of water,nutrients, vitamins, minerals, amino acids, proteins, oils, fatty acids,saccharides and polysaccharides. Preferably methods of applying comprisespraying or misting and the area is a walking path, a mining area,tailings, gangue, a debris or waste pile, a cliff, a roadway, anaircraft landing area, a contaminated or hazardous area, or aconstruction site, or air space of a mining area, factory, manufacturingplant, or work area. Preferably, either or both compositions form a filmon a surface that promotes binding of the microorganisms and may beaqueous. Also, the film may comprise a polymer, and either or bothcompositions may contain a coloring agent, or a detectable marker.Applying may comprise repeated applications of either or bothcompositions to the surface, and/or further applying another aqueouscomposition to the area.

Another embodiment of comprises kits for dust control comprising: afirst composition containing a mixture of viable cells or spores ofurease-producing microorganisms and a transport media; and a secondcomposition containing a form of calcium, wherein either or both firstand second compositions further contain urea and ingredients for growthof the mixture of viable cells or spores. Preferably the mixture ofviable cells or spores comprise yeast, algae, bacteria, eukaryoticcells, or recombinantly engineered microorganisms, and preferably thebacteria comprise Sporosarcina pasteurii, Bacillus megaterium,Sporosarcina ureae, Proteus vulgaris, Bacillus sphaericus, Myxococcusxanthus, Proteus mirabilis, Helicobacter pylori, or variants, serotypes,mutations, or combinations thereof. Preferably the transport mediummaintains viability without promoting significant propagation of themicroorganisms, such as minimal cell medium. The form of calcium maycomprise calcium chloride, calcium acetate, calcium phosphate, calciumcarbonate, calcium lactate, calcium nitrate, and/or a calcium salt, andthe ingredients for growth of microorganisms comprise one or more ofwater, nutrients, vitamins, minerals, amino acids, proteins, oils, fattyacids, saccharides and polysaccharides. Preferably either or both firstand second compositions are aqueous, may further contain a polymer, acoloring agent, and/or a detectable marker. Kits may also comprise anapparatus for applying either or both first and second compositions.

Other embodiments and advantages of the invention are set forth in partin the description, which follows, and in part, may be obvious from thisdescription, or may be learned from the practice of the invention.

DESCRIPTION OF THE INVENTION

Traditional constructions materials such as clay bricks and concreterequire enormous amounts of energy during the manufacture process. Theseprocesses are heavily reliant on burning natural resources such as oil,coal and wood. This reliance results in the consumption of massiveamounts of energy resources and equally massive carbon dioxideemissions, thus a great dependency on limited energy sources. Analternative has been described which requires much less energy formanufacturing that utilizes enzymes produced by microbial cells.Typically, cells are aerobic and/or facultative anaerobic cells andinclude, for example, Sporosarcina pasteurii, Bacillus megaterium,Sporosarcina ureae, Proteus vulgaris, Bacillus sphaericus, Myxococcusxanthus, Proteus mirabilis, Helicobacter pylori and other strains,serotypes, variations, mutations and CRISPR modifications (clusteredregularly interspaced short palindromic repeats). Cells produce theenzyme urease which, in the presence of calcium and nitrogen sources,forms calcite crystals. The process is generally referred to a microbialinduced calcite precipitation (MICP), which can be performed with thecells or purified enzyme. As little to no heating is necessary, theenergy savings in both expenses and efficiency is enormous.

The enzymes and/or the enzyme-producing cells are dispersed in acomposition containing a nitrogen source and a calcium source, such asfor example, urea, and calcium chloride with an aggregate materialcatalyzing the production of ammonia and carbon dioxide, increasing thepH level of the composition. The rise in pH forms a mineral precipitatecombining calcium with carbon dioxide. The cells or other particles actas nucleation sites, attracting mineral ions from the calcium to theparticle surfaces forming calcium carbonate crystals such as calcitecrystals or other calcium carbonate polymorphs. The mineral growth fillsgaps between the particles of aggregate, bio-cementing or bondingaggregate particles forming a solid. The resulting material exhibits acomposition and physical properties similar to naturally formedsandstone, but whose hardness can be predetermined based at least on thestructure of the initial components and the pore size desired.

Compositions and methods have been surprisingly discovered that areuseful in the control of dust. The compositions include two solutions, afirst solution containing a fluid plus urease enzyme or urease-producingcells or cell spores and a second aqueous solution containing calciumions. The first or second solution may contain nutrients for thepropagation of the cells or the spores, a nitrogen source such as,preferably urea, and a carrier such as clay or pond fines. Commercialsources of urease enzyme include, for example, jack beans. The enzymescan be maintained as a liquid, but are preferably lyophilized for easeof storage and transport, and re-hydrated before use with a fluid suchas, preferably, water, buffered water, or another hydrating agent thatpreserves enzyme activity. Preferable, pure enzyme is encapsulated incarbohydrate, lipid or other polymer microshells or spheres.Encapsulation techniques include, for example, encapsulation innanoorganized microshells, and encapsulation in xanthan-alginatespheres. Preferred enzyme concentrations are from 0.5-5 mg/ml in 0.1 Mphosphate buffer, pH 7.6. Preferably enzyme concentrations are fromabout 0.1 to 100 mg/ml, more preferably about 0.5 to 3.0 mg/ml, morepreferably from about 0.5 to 2.0 mg/ml, and more preferably about 1.0mg/ml. Enzyme can be further diluted prior to use to obtain a rate of0.02-0.04 AA/minute. Enzyme activity can be measured by the reaction:

which couples ammonia production to a glutamate dehydrogenase reaction.Accordingly, one unit of enzyme results in the oxidation of onemicromole of NADH per minute at 25° C. and pH 7.6.

This method for manufacturing construction materials through inducedcementation exhibits low embodied energy and can occur at ambientpressures or higher or lower, and ambient temperatures or higher orlower. For example, preferred pressures are from about 10 psi to about100 psi and all pressure values in between, also preferred at from about14 psi to about 50 psi. Although higher pressures can be utilized, thereis generally no need for the energy expense required. Preferredtemperature ranges are from at least minus 20° C. to above 80° C.,preferably from about 5° C. to about 50° C., preferably from about 15°C. to about 30° C., preferably from about 20° C. to about 25° C.Preferably, temperature ranges are below 30° C., below 40° C., below 50°C., below 60° C., or below 70° C. The ambient temperatures andconditions as well as the content of available aggregates can determinewhether pure enzyme, lyophilized enzyme, spores, or live cells areutilized as the starting components. Living cells can be used intemperatures where mild weather conditions exist, whereas pure enzymescan be advantageous at more extreme conditions of cold or heat. Sporesare used when immediate calcification is not required and the spores areprovided sufficient time to germinate and express enzyme.

Processing also involves the production of quantities of by-productssuch as ammonia, not all of which is utilized in calcite formation. Itis another embodiment of the invention to include an effluent recoverysystem in association with the production methodology. The recovery ofammonia from effluent converts the effluent to plain water which can berecycled or disposed of without any need for additional decontaminationprocedures. Preferred ammonia recovery methodologies include, forexample, ion-exchange resins and commercially available processing suchas Ammonia Electrolysis, zeolite, clinoptilolite and combinationsthereof. Preferably, the ammonia recovered can be utilized infertilizers, converted to nitrogen, utilized for energy generation orutilized for other applications.

One embodiment of the invention is directed to compositions comprisingurease-producing cells or cell spores, urease enzymes (e.g., crudeextract, or unpurified or purified enzymes) in a transport medium andoptionally a nutrient medium. Transport medium includes, for example,growth media for urease-producing and/or other supporting cells, enzymestabilizing media, reagent media, buffered solutions and combinationsthereof. The composition may include or be combined with a supportmaterial which may be organic or inorganic and is preferably a solid orsemi-solid and preferably contains holes or perforations and/or isotherwise porous. Organic support material includes, for example,biomass such as, preferably, moss, hay, straw, grass, sticks, leaves,algae, dirt, ash, dust, particulate material, refuse and combinationsthereof. Inorganic material includes, for example, minerals,supplemental cementitious materials (SCM), pulverized or crushed rock,fines, and combinations thereof. Fibrous materials include sheets ortarps of burlap, paper, wood (e.g. residuals), cotton, or anothernatural or synthetic fiber. Non-natural and manufactured materials mayalso be used such as, for example, sheets of plastic, glass, fiberglass,vinyl, rubber, synthetic fabrics or combinations thereof. To the solidsupport is applied or otherwise introduced urease-producing cells,urease enzyme or simply other cells. Preferably these other cells wouldbe useful to support the grown of the urease-producing cells or enhancethe chemical processes involved and not otherwise interfere with theMICP process or to act as nucleation sites. Preferably these other cellsare native or latent microorganisms in the local environment or providedwith the mix, non-pathogenic, non-toxic and/or relatively harmless atthe amount used, and easily obtained, present in the local environmentor provided. Cells can be proliferated directly on the support materialand, at a desired density or growth stage, the organic material evenlydispersed and/or thoroughly mixed into an aggregate material formanufacture of construction tools and products. Inorganic materials thatcan be used include, for example, rock (e.g., fines), sand, glass, wood,paper, metal, plastic, polymers, minerals, manufacturing or processingwaste materials such as ash, carbon or wood residuals, any of which canbe crushed or used whole or combinations thereof. Compositions may alsobe formed from waste materials that are otherwise hazardous (e.g.,radioactive materials, materials with dangerous metal or poison content,contaminants from scrubbers, or other harmful materials) and formed intosolid structures that can be stably stored or otherwise safely disposed.

Compositions of the invention may be sprayed or otherwise applied tosheets or mats or natural or non-natural materials and the sheets used,for example, to prevent erosion by formation of a calcium carbonatecrust over a surface, a pile, a cliff or other structure that's subjectto erosion. With the use of perforated or porous sheets or mats, thecrust forms through the support material attaching the material on whichthe sheet has been placed. Nucleation sites for calcite formation caninclude, for example, polymers, fibers, fines, SCM, added Portlandcement, powders, co-cultivated microorganisms, and combinations thereof.One or multiples layers of crusts can be formed on site. In this way,erosion and dust control can be substantially reduced or eliminated indefined areas. Importantly, in this fashion the sheet can be easilyreplaced over time and/or a fresh composition of the inventionre-applied to the surface as needed or as desired. Mats provide theadditional benefit of “seeding” the site for rehabilitation onceoperations have ceased—thereby allowing site recovery efforts to proceedsuch as returning the site to a natural state. This is especiallyapplied to mining sites in which mining operations have ceased.Preferred areas for application include a walking path, a mining area,tailings, gangue, a debris or waste pile, a cliff, a roadway, anaircraft landing area, a contaminated or hazardous area, or aconstruction site, or air space of a mining area, factory, manufacturingplant, or work area.

In a preferred embodiment of the invention, compositions of theinvention are applied to a surface area as a liquid, a gel, a slurry, asludge, a semisolid or a dry powder. Spores and/or microorganisms of acomposition of the invention produces enzymes that catalyze formation ofa crust of calcium carbonate in the presence of liquid, which ispreferably water, buffered water or another aqueous material. A nutrientmix suitable for the particular microorganism can be included with thecells. When the composition dries, the crust remains and cells godormant. As cells self-propagate, provided sufficient nutrients and/orsubstrate materials are present, new crust will form whenever sufficientaqueous liquid is provided. In a preferred embodiment, nutrients and/orsubstrate materials may be distributed over the surface area of interestin slow-release or timed-release form such as dry components with apre-determined rate of dissolution. Reformation may simply be a matterof re-applying water that dissolves nutrient and/or substrate therebyre-activating the microorganisms. The re-activated microorganismsproduce enzyme that forms crust. This process can be repeated with orwithout the reapplication of microorganisms, nutrients and/orsubstrates, or with only occasional additions. This process may becoupled with weather events so that the rain provides the source ofwater. By providing a composition that provides microorganisms and/orspores and contains slow-release nutrients and/or substrate, crust canbe reformed over an area repeatedly over long periods of time.

Preferably compositions of the invention including all necessarycomponents such as microorganism, spores and/or enzymes, a nitrogensource, and a calcium source and optionally nucleation sites (e.g.,powders, fines, co-cultured microorganisms and/or other materials), areapplied to a surface, such as, for example, a dirt road, or a structuresuch as a hill or cliff. Microorganisms proliferate and produce enzymewhich h catalyzes the formation of a crust of calcium carbonate over theroad surface. As vehicles travel over the road, the crust breakseventually turning the crust into dust. Periodically the geographicregion experiences rain or other forms of precipitation that dissolvesslow-release nutrients and/or substrate materials, thereby promotingproliferation of dormant microorganisms. The microorganisms produceenzymes which catalyze new crust formation over the road. In periods ofreduced precipitation, an aqueous solution is re-applied to the road toactivate the microorganisms which may or may not contain additionalnutrients and/or substrate materials. Preferably the composition is aliquid, a gel, a slurry, a sludge or dry powder and the support materialmay be in the form of beads, grains, rods, strands, fibers, flakes,dirt, biomass, sand, pulverized or crushed stone, fines, supplementalcementitious materials (SCM), crystals, co-cultivated microorganisms, orcombinations thereof. Fines sizes are preferably equal to or less than250 micron, more preferably equal to or less than 200 micron, morepreferably equal to or less than 150 micron, or more preferably equal toor less than 100 micron (reference examples include micron size of beachsand=700, micron size of fine sand=250; micron size of Portlandcement=74; micron size of silt=44; micron size of smoke=2). Supportmaterial and aggregate material can be the same or different. Preferablythe support or aggregate material is sand, glass, metal, added Portlandcement, SCM, fines, co-cultivated microorganisms (e.g., native, latent,local, added, or genetically modified microorganisms), or combinationsthereof and the urease-producing cells or cell spores comprise yeast,algae, anaerobic cells, facultative anaerobic cells, bacteria oreukaryotic cells or cell spores. Preferred bacteria are Sporosarcinapasteurii, Sporosarcina ureae, Proteus vulgaris, Bacillus sphaericus,Myxococcus xanthus, Proteus mirabilis, Helicobacter pylori, or variants,serotypes, mutations or combinations thereof, and preferred yeast,algae, bacteria or eukaryotic cells or cell spores are geneticallyengineered. Other enzyme producing bacteria that are capable ofbiocementation include Sporosarcina ureae, Proteus vulgaris, Bacillussphaericus, Myxococcus xanthus, Proteus mirabilis and Helicobacterpylori, although proper concerns should be given to pathogenic strains.Combinations of any of these strains as well as functional variants,mutations and genetically modified stains may be used as well. Thesupport material and the cells are preferably coupled via hydrophobicbonds, hydropyllic bonds, ionic bonds, non-ionic bonds, covalent bonds,van der Waal forces, or a combination thereof and/or the supportmaterial is at least partially or totally encompassed by a film thatpromotes binding of the urease-producing cells. Preferred films comprisea polymer or a cell nutrient and preferably the composition contains acoloring agent which may be red, blue, green, yellow or any combinationor shade thereof. Preferably the composition contains an identifyingagent or a detectable marker such as a microscopic tag, a color, anenzyme or another substance.

Support materials and/or aggregate materials may contain additionalcomponents that provide an advantage to the construction materials. Forexample, chemicals and/or additional cells (e.g., native, local orlatent bacteria, yeast, eukaryotic cell, algae, and recombinantvariations thereof), can be included that produce enzymes, cofactorsand/or other chemicals useful in breaking down stains in and/or acquiredby the final product and/or additional nucleation sites. Stains includestains from air pollution, soot, mold or animal waste products.Alternatively, the chemical or enzymes may impart color, texture or adesired function or appearance to the final product.

Another embodiment of the invention is directed to kits formanufacturing solid forms comprising: the composition of the invention,a second composition containing nutrients for proliferation of theureases-producing cells and/or germination of the cell spores; aplurality of sets of formworks wherein each set encloses the shape of atleast one solid form and contains one or more porous panels; and a thirdcomposition comprising a calcium source (e.g., CaCl₂), a nitrogen source(e.g., urea) or both a calcium source and a nitrogen source. Preferablythe kit is for the creation of solid forms such as, for example,rectangular, square, rounded, oval or an irregular shape. Preferredsolid forms include but are not limited to blocks, boards, bricks,pavers, panels, tiles, counter tops, or veneer. Preferably kits of theinvention are for the manufacture of blocks such as, for example,concrete masonry, cinder blocks, foundation blocks, breeze blocks,hollow blocks, solid blocks, besser blocks, clinker blocks, high or lowdensity blocks, or aerated blocks, thin bricks, manufactured stone,cultured or colored stone. Nutrient compositions of the invention maycontain nutrient media to maintain and/or allow the cells to flourishand proliferate. The various types of nutrient media for cells, and inparticular, bacterial cells of the invention are known and commerciallyavailable and include at least minimal media (or transport media)typically used for transport to maintain viability without propagation,and yeast extract, molasses, and corn steep liquor, typically used forgrowth and propagation. Preferably the nutrients include amino acids,proteins, polysaccharides, fatty acids, vitamins and minerals.

Another embodiment of the invention is directed to methods formanufacture of solid forms comprising: mixing the composition of theinvention with an aggregate material and water to form a mixture,wherein the aggregate material is largely composed of particulates ofless than 5 mm in diameter (e.g. less than or about 4 mm, less than orabout 3 mm, less than or about 2 mm, or less than or about 1 mm);apportioning the mixture into multiple form works wherein each form workcontains at least one porous panel; adding a second composition to themixture, wherein the second composition contains nutrients that promoteproliferation of the urease-producing cells; adding a third compositionto the mixture, wherein the third composition is a liquid, powder orpaste that contains calcium; incubating the mixture for a period of timeto form covalent bonds between the particulates; and removing the solidforms from the form works. Alternatively, the compositions may becombined and added together to the material within the form works orcombined with the material prior to addition to the form works.

Another embodiment of the invention is directed to the structure andcomposition of form works. Preferred form works comprises athermoplastic material that can be molded or extruded into a desiredshape. Preferred thermoplastics include, but are not limited to plasticssuch as polypropylene, polystyrene, polyethylene including HDPE (highdensity polyethylene), LPDE and reclaimed LDPE (low densitypolyethylene), and cross-linked polyethylene, glass and most anyformable polymer. Preferably, the polymer material is provided aspellets or lens shapes that range in thickness and uniformity. Thepellets are filled in a porous mold and steamed under pressure (the moldis not under pressure, pressure just from the steam). The resultingproduct provides a designed flow directional material, and changes tothe gradation impact the flow direction, speed and retained saturation.

Another embodiment of the invention is directed to compositions andstructures that do not require formworks (e.g. frameless manufacturing)wherein structures are formed from a combination of the components ofthe invention plus polymers and/or thermoplastics that are compressedwith a compaction device and retain the desired structure. Preferredcompression devices include hydraulic presses and preferred pressuresare 100 psi or greater, 250 psi or greater, 500 psi or greater, 1000 psior greater, 2000 psi or greater, 3000 psi or greater, 4000 psi orgreater, 5000 psi or greater, Preferred components of the inventioninclude all the components to form calcium carbonate structures in theform of a sludge or paste. The compaction device compresses thecomponents with added pressure into a form that is maintained and drieswithout significant alterations of the resulting form. Preferredpolymers and thermoplastics include, but are not limited to plasticssuch as polypropylene, polystyrene, polyethylene including HDPE (highdensity polyethylene), LPDE and reclaimed LDPE (low densitypolyethylene), and cross-linked polyethylene, glass, carbohydrates suchas starches, lignin, and most any formable polymer. Compressed form canbe generated rapidly from a think slurry or sludge and maintains itsshape during calcite formation. Preferably calcite formation isaccomplished in vapor chambers (e.g., at greater than ambient pressures)that contain increased vapor pressures or are sprayed or misted, whereinthe vapor, mist or spray preferably comprises nutrients or chemicalsubstrates. Preferred forms include blocks, bricks, thin bricks,manufactured or cultured stone, pavers, or any useful structure.

Preferably the multiple form works or compression devices create 5, 10,50, 100, 500, 1,000, 10,000, 100,000 1,000,000 or more forms at a time.The number of form works or compression devices that can besimultaneously utilized is limited only by the complexity of themechanics and space available. These form works or devices may bestacked or provided in a single layer or pallet. Formwork may havevertical walls which are connected together forming cavity there betweento receive the aggregate material. Formworks may also have a floor and,alternatively, the bottom of the formwork may be left open if supportedby a porous surface such as soil, or aggregate and composition may bemixed and pressed into molds or extruded. Preferably, vertical walls areat least the inside surfaces thereof, are made of a non-reactive,non-porous material or coating such as cast or extruded acrylic resin.This enables one to easily remove the construction material or the brickfrom the formwork after it has solidified. In addition, the verticalwalls and floor of formwork or pressure devices may have designs thatform surface textures in the resulting bricks or other structures (e.g.,lines, circles, waves, groves, sketches, images, etc.).

Preferably the aggregate material comprises rock, glass, fiberglass,wood (residuals, pulp, sawdust, lignin), biomass, paper, metal, plastic,polymers, rubber, imitation rubber, vinyl, minerals, co-culturedmicroorganisms, waste materials (e.g., ash, carbon, scrubber waste,radioactive pellets) or combinations thereof, and/or mixing comprisingspraying the composition as a liquid onto the aggregate material.Preferably the form works are substantially submerged during theincubating and air is bubbled to the submerged form works. Preferably athird composition is added to the mixture repeatedly during incubatingwhich drains through the bottom panel and, optionally, is recycled.Preferably, incubating is performed under ambient conditions and thethird composition contains calcium chloride, calcium acetate, calciumphosphate, calcium carbonate, calcium lactate, calcium nitrate, or acalcium salt. Preferably the pH of the mixture is monitored during theincubating. Preferably the solid forms are blocks, boards, bricks,pavers, panels, tiles, or veneer, and the mixture further containsfibers or nanofibers that are, for example, fibers or nanofibers ofwood, glass, plastic, metal or a polymer. The solid forms can bepartially or uniformly porous containing a network of holes or gaps.Holes can be of a predetermined size and/or structure such as, forexample, at least 5 microns, at least 10 microns, at least 20 microns,or at least 50 microns in diameter. Alternatively, solid forms can bemanufactured with materials that provide virtually no or few holes. Forexample, adding a non-porous material to the aggregate mixture cancreate complex and extended pathways that render the form impermeable tofluids.

Another embodiment of the invention comprises compositions containingurease producing cells or cell spores that are coated with nutrientmedia. Preferably the nutrient media further contains additional ureaseproducing cells or cell spores, and/or nutrients to promote theproliferation of additional cells that have been added to the aggregatethat are beneficial to the final product.

Another embodiment of the invention is directed to compositions, methodsand systems for the treatment of aggregate materials comprised ofparticles with a composition comprising one or more of a nitrogen sourcesuch as for example urea, a calcium source (e.g., calcium ions) andurease or urease producing cells. Preferably particles have a diameter(e.g., actual, average or effective diameter) of about 50 mm or less,preferably about 25 mm or less, preferably about 20 mm or less,preferably about 10 mm or less, and preferably about 5 mm or less. Inone preferred embodiment, aggregate material can also be about 1 mm orless and preferably about 0.5 mm or less, more preferably about 0.1 mmor less, and more preferably about 50 μm or less. Especially preferredparticles sizes include from about 10 μm to about 1 mm, from about 100μm to about 0.5 mm, from about 200 μm to about 1 mm, from about 1 μm toabout 200 μm, from about 10 nm to about 1 μm, and from about 10 nm toabout 40 nm, and various combinations thereof. Particles include, forexample, spores, carbon dust, dust or soot from cement or brickmanufacture, cement block manufacture, foundry operations, grindinglimestone, sand tailings, mining, smelters, paint manufacturing andbyproducts of other manufacturing processes such as slag. Particles maybe obtained and collected from available or implemented dust controlprocedures. Particles may be of mixed sizes including but not limited tosizes equal to and greater than preferred sizes, particles equal to andless that preferred sizes, and combinations of preferred sizes andmixtures thereof. Particles that are aggregates and more sizableparticles may include recycled and/or recyclable materials. The nitrogensource of the composition may be a single chemical, such as urea of anygrade and purity and is preferably commercially obtained. Calcium ionsare preferably obtained from commercially available sources such as, forexample, calcium chloride. Urease enzyme or urease-producing bacteriamay be included in the composition. Urease-producing bacteria include,but are not limited to the bacteria Sporosarcina pasteurii, Sporosarcinaureae, Proteus vulgaris, Bacillus sphaericus, Myxococcus xanthus,Proteus mirabilis, Helicobacter pylori and combinations thereof. Ureaseproducing cells includes non-viable cells that contain enzyme such as,for example, mycells, cells composed of lipids or fatty acids, and cellscontaining urease. Urease and/or urease-producing cells may produce orrelease a predetermined amount of enzyme over a defined period of time.Preferably, the amount of urease released per cell is sufficiently rapidto allow for the rapid creation of calcium carbonate in the presence ofnitrogen and calcium ions.

Preferably, particles are combined with a nitrogen source (e.g., urea),urease and/or urease producing cells, calcium ions and preferably waterto create a homogenous slurry. The slurry can be painted or sprayed ontoobjects and/or surfaces creating a layer or crust, molded into formsthat solidify into objects which may be complete or partially solid, orotherwise pooled for immersion or dipping of objects to be coated withthe slurry material again creating layers or a crust over the objectsurfaces. Objects may contain one or more layers as desired, and layersmay be permeable or impermeable to water or improve resistant to wearfrom weather conditions such as sun damage, snow, ice and rain. Slurriesthat provide increased resistance are preferably composed with aggregatematerials that are particles of less than 0.1 mm diameter. As the liquiddries, calcium carbonate bonds form between the particles and/or theparticles and the object. The result can be an object containing anouter shell of hardened calcium carbonite or a formed structure. Objectsthat can be manufactured according to the invention and/or layered witha crust or coating of the invention include, but are not limited tobricks, cement blocks, pavers, counter tops, glass, fiberglass, polymerand acrylic structures, siding, walls, yard art, slate and rockstructures, tiles, paving stones, steps, roofing material, gutters,cement walls and planks, patios, balconies, fencing and combinationsthereof.

Another preferred embodiment of the invention comprises producingammonia and/or other compounds (e.g., ammonia, organic acids, alcohols,phenolics, sulfides) by fermentation of microorganisms (e.g.,microorganisms that produce ammonia monooxygenase, hydroxylamineoxidoreductase, nitrifying bacteria). Preferably microorganisms areselectively cultured to maximize generation of the desired enzymes.Hyper-ammonia producing microorganisms include, for example,ruminant-derived microorganisms, intestinal microorganisms,Peptostreptococcus sp., Clostridium sp., Calliandra sp., Atopobium sp.,Desulfomonas sp., and the like. Isolated ammonia can be recycled orutilized in other processes such as in fertilizers and energyproduction.

Another embodiment of the invention comprises spraying the slurry of theinvention onto a natural geological or man-made surface such as a cliff,a dune, an aggregate pile, a ledge, a supporting wall, ores, afoundation, minings, tailings, piles of waste materials from amanufacturing process, construction site, or another structure for whichadditional support or structuring is desired. Such support isadvantageous in convenience and financial considerations as compared toproviding additional support of the structure of interest withconvention building systems. In addition, and preferably, slurry of theinvention can be provided to geological surfaces such as soil aroundbuildings to provide building support, erosion suppression, preventionand/or repair of sink holes, or to create foundation structures toprovide solid support and/or stabilization of buildings and otherstructures, and combinations thereof.

Another embodiment of the invention is directed to compositions, methodsand systems comprising a slurry of one or more of water, a nitrogensource (e.g., urea), optionally a calcium source (e.g., CaCl₂), andurease or urease-producing cells, but without the addition or anyaggregate material such as, for example, without sand, soil, dust, siltor other particles as aggregate materials. Preferably the slurrycontains at least water, a nitrogen source (e.g., urea), optionally acalcium source (e.g. CaCl₂), and urease or urease-producing microbes,which may include microbial nutrients as appropriate. Preferably thecalcium source is provided is another composition and the slurry iscalcium-free. This liquid slurry is sprayed, painted, or otherwiseplaced directly on or in an aggregate material, or formed in a mold ofmost any shape or structure containing aggregate material, optionallyfollowed by application of the calcium composition. The combination ofthe aggregate and the aforementioned slurry forms a solidified object,covering or layer (or multiple layers), such as, for example, as abuilding foundation, a molded object, a layer covering an object, oranother desired form. One of the advantages of this technique is thataggregate material does not need to be shipped and there is aconcomitant associated savings. Preferably aggregate material isimmediately available on site or locally available within an acceptabledistance. The addition of the slurry to the already-present aggregatecreates a solid or more hardened form of a structure efficiently insituations where transporting or otherwise moving aggregate materialswould be difficult, inefficient or impractical such as but are notlimited to situations involving creation of, repair of or to furthersupport building foundations and other repairs.

Another embodiment of the invention is directed to compositions andmethod comprising a slurry of the invention combined with an aggregatematerial, further containing multiple solid structures that are eitherhollow or otherwise of lighter weight than the aggregate material. Theresulting structure containing the additional objects produce solidobjects of lighter weight than objects made of only aggregate materialand slurry. Alternatively, it may be desirable to increase the weight ofthe object by adding objects that are heavier than the aggregatematerial. Such heavier objects include, but are not limited to rebar orremesh, metal forms, strengthening material and other heavier materials.These additional objects include, but are not limited to plastic, wood,steel, metal, polymer, rods, balls geometric structures, which may besolid, perforated or hollow. Alternatively, the additional objects maybe included that have aesthetic properties such as, for example,predetermined colors, materials, functions, properties and designs. Thisis advantageous when light weight objects are desired, wherein thestructure retain sufficient strength for the intended purpose such as,for example, a specific desired compression strength, tensile strength,yield strength, ultimate strength, Young's modulus, elastic modulus,elastic strength, stiffness, hardness, toughness, stress resistance, andcombinations thereof.

Another embodiment of the invention is directed to compositions, methodsand systems comprising a variety of substrates combined with a slurry ofthe invention. The addition of sand, fines, silt, or dust (which arelighter and have smaller particles than soil or other aggregates) tourea, urease, optionally calcium and water create lighter structureswith equivalent or nearly equivalent support strengths. Preferably theslurry is largely calcium free and a calcium-containing composition isprovided in a second application. Advantages of lighter structuresinclude a lower cost of production and a higher efficiency ofproduction, as well as other benefits such as efficiency or manufactureand formation of structures. Preferably, urease enzymes are used toincrease the solidification of the structure as compared to the use ofenzyme-producing cells. In addition, enzymes are smaller in molecularstructure than cells and will pass through smaller pore sizes ofaggregate materials having small pore sizes. Also in addition, one ormore chemical or compounds can be included to increase the densityand/or weight of the liquid composition so that compositions settlesquickly or are sufficiently sticky to a surface (e.g., as a gel, foam orsemi-solid).

Another preferred embodiment of the invention comprises composition,systems and methods for forming solid or porous solid structuresaccording to the invention that are lighter in weight as compared toconvention structures composed of clay or cement. Preferably, theinvention comprises creating a spatial gap within the solid structureduring manufacture as the structure hardens. This gap can be in the formof holes, tubes, bubbles, or any other three-dimensional shape. Apre-formed shape made of the same aggregate material or materials, or ofa different, preferably lighter material can be immersed into the wet,un-hardened slurry of the invention either with or without aggregatematerial. When the slurry fully formed around that desired shape, theresulting object will weigh less than conventionally prepared objects,such as, for example, clay bricks, cement blocks, pavers, stonecomposites, or another solid structure composed of one or more aggregatematerials. The resulting solid object has an increased strength, new orenhanced aesthetic or performance characteristic, additive or acombination thereof.

Another preferred embodiment of the invention comprises composition,systems and methods for forming protective layers or coverings to solidstructures. Preferably the slurry of the invention fills and closespores in the solid structure (e.g., a fabric impregnated with one ormore of microorganisms, nutrients, substrate materials, nucleationsites) so as to provide effective barriers to liquids (e.g. water),gasses (e.g. pollution) or other substances that may impregnate orcontaminate a solid structure. Such compositions can be used for erosioncontrol and structural support.

Another embodiment of the invention comprises compositions, systems andmethod for dust control of, for example, walking paths, piles, aircraftrunway, taxi ways, and parking areas, cliffs, vehicle roadways and otherlarge surfaces. Slurries of the invention can be substituted for oilsand other dust control compositions presently used on dirt, gravel andother road surfaces to minimize the amount of dust created fromvehicles. Slurries of the invention can be sprayed or vaporized fromtrucks or other vehicles as a liquid, or administered (e.g., spraying)as a dry composition to be activated when wetted, onto surfaces forminga hardened crust to the road or other surface. Slurries comprisingsubstrates and living urease-producing microbes plus nutrients coverroad surfaces with a self-renewable crust. An initial application caninclude microorganisms and optionally included with subsequentapplications which may only contain substrate materials. As vehiclespass over the road, the crust may be damaged from the weight of thevehicle, but a crust is recreated and repaired by the presence of theliving-slurry. Preferably slurry of the invention for dust controlcontains no aggregate or only aggregate of 0.5 mm or less in diameter.

Another embodiment of the invention comprises adding slurry of theinvention, either with or without aggregate, to conventional proceduresfor the manufacture of construction materials such as, for example, claybrick, cement blocks, pavers, and other substances. Slurry additions canbe included as desired at from 0.0001 percent to 99 percent of the dryweight of the resulting product or empirically determined from the typeof aggregate used. Preferably the slurry addition by dry weight is from1 to 50 percent, from 2 to 75 percent, from 30 to 60 percent, from 25 to80 percent, from 10 to 25 percent or any combination there.

Another embodiment of the invention comprises creation of a slurry ofthe invention with which will solidify at a predetermined time.Preferably, slurries contain a predetermined amount of nitrogen andcalcium sources as substrates and a predetermined amount of enzyme thatsolidifies within a desired time frame. Solidification conditions mayinclude the temperature of use, which can be included in thecalculations to determine solidification times preferably experimentallyor empirically.

The following examples illustrate embodiments of the invention, butshould not be viewed as limiting the scope of the invention.

EXAMPLES Example 1

Reducing dust of surface mining sites is required by MSHA (miningversion of OSHA) regulations. Current methods used in the industryinclude the use of various polymers or chemicals, with the most commonbeing the continuous spray application of water, oils and other dustcontrol liquids. The objective for surface mine dust control is to makefine dust (a byproduct of aggregate mining) heavier than air to preventrespiratory and visibility hazards. According to the invention,micro-organisms are applied either with a nutrient and/or transportmaterial or in association with any conventional treatment for such dustcontrol and/or surface cementation and include production of a calcitecement (CaCO₃) in combination with urea (nitrogen/carbon) and a calciumsource. Cells and/or nutrient materials are included with an initialapplication, and optionally with subsequent or follow-on applications.Preferably, applications are of light-weight materials that are quick tocement using the same strains of urease-producing bacteria as used inthe formation of bricks, pavers and other solid forms. Alternatively,cyanobacteria, a photosynthetic microorganism that fixes nitrogen fromthe atmosphere, is substituted for or used in addition tourease-producing bacteria, which reduces nutrient input needs.

Example 2

Recovery systems seek to address: (a) returning effluent to a viablestate to be re-used as influent (water becomes a capital expense ratherthan a consumable material), and (b) the extraction of commerciallyvaluable byproducts from the effluent stream. Preferably, thebio-cementation process of the invention is useful for the primaryproduction of by-products as products such as, for example, using ureaseproducing microorganisms for the manufacture of ammonia/ammonium and/orfree calcite. By-products are excess material to be reduced throughoptimization and/or accounted for in influent formulations. Ammonia as arecoverable by-product has commercial value in both fertilizer andalternative fuel applications.

There are at least two ammonia extraction methods. First, granularzeolite cliniptilolite mineral aggregate is used as an air filter forextracting ammonia gas, and as a liquid filter for extracting ammoniumfrom effluent. Ammonia-saturated zeolite has potential applications as afertilizer, fertilizer additive, and/or fertilizer component. Second, anelectrode based system is used for conversion of aqueousammonia/ammonium as a hydrogen fuel source for electricity production.

Effluent, further treated or not, is a fuel source for otherammonia-based energy production technologies and recycling technologiesincluding recycling of water, calcite and by-products.

Settling tanks, mesh filters, fabrics and/or hydrocyclones are used forthe removal of free calcite in solution wherein, and preferably, themicro-organisms remain. This material is an inoculation source for newbiocement formation and fertilizer applications (calcium available forplant cell wall formation, and microbes available for soildenitrification).

Example 3

Biologically-formed Microcrystalline Calcium Carbonate was producedusing a urease-producing microorganism (S. pasteurii) grown in a liquidfermentation medium containing urea. The media was agitated to create auniform suspension. At late stage growth of the culture, calcium ionswere added in the form of a calcium chloride solution to a saturation ofmolar equivalency with the urea. Urease activity results in thehydrolysis of urea (2NH₂CO) into ammonium (NH₄) and carbon (C). Carboncombines with calcium (Ca) to produce calcium carbonate (CaCO₃). Calciumcarbonate crystals formed ranged in size from 50 μm to 0.1 μm andgenerally “regular” (e.g., spherical) in shape. Calcium carbonate wasseparated from the solution using one or more of centrifugation,settling tanks, hydro-cyclones, or decanting. The method was performedas a batch process and also as a continuous production line.

A variation of this method was used to increase particle size by usingfine aggregate materials to create an agglomerate, which also improvedliquid-solid separation. In this variation, fine aggregate with a MeshScale of 70 micron size was added to the solution during thefermentation process and at a quantity that did not exceed the abilityfor the agitation to keep the fine aggregates in suspension. Followingthe addition of calcium ions, the calcite bonds to, and bonds togetherthe fine aggregate, creating larger, heavier particles.

Example 4

The method of Example 3 is performed with co-culture of a secondorganism, Delaya venusta. The co-culture process is developed for asingle fermentation within a single reactor, alternating fermentationswithin a single reactor, or by circulating media between two separatedfermentations in separate reactors, wherein the reactor is either aliquid state reactor (e.g., batch, batch-fed, or continuous), or asolid-state reactor such as an aggregate-unit (e.g., bricks).

Example 5

In another example, a first composition is prepared containing anaqueous mixture of viable cells or spores of urease-producingmicroorganisms suspended within a transport media that promotesviability and not growth and/or germination of spores, but with minimalor no calcium. A second composition is prepared that contains an aqueousform of calcium, calcium chloride. A third composition is preparedcontaining urea and ingredients for growth of the microorganisms or,alternatively, the urea is added to the first composition and theingredients are added to the second composition. The compositions arethan applied to an area simultaneously or sequentially and, ifsequentially, in any order. To an area such as a mining operation, dustin the atmosphere is significantly reduced and mostly eliminated.

Other embodiments and uses of the invention will be apparent to thoseskilled in the art from consideration of the specification and practiceof the invention disclosed herein. All references cited herein,including all publications, U.S. and foreign patents and patentapplications, are specifically and entirely incorporated by reference.The term comprising, where ever used, is intended to include the termsconsisting and consisting essentially of. Furthermore, the termscomprising, including, and containing are not intended to be limiting.It is intended that the specification and examples be consideredexemplary only with the true scope and spirit of the invention indicatedby the following claims.

1. A method of dust control comprising: providing two compositions,wherein one composition contains an aqueous mixture of viable cells orspores of urease-producing microorganisms and a transport media, and theother composition contains an aqueous form of calcium, wherein either orboth compositions further contain urea and ingredients for growth of themicroorganisms; and separately applying the two compositions to an area.2. The method of claim 1, wherein the microorganisms comprise yeast,algae, bacteria, eukaryotic cells, or recombinantly engineeredmicroorganisms.
 3. The method of claim 2, wherein the bacteria compriseSporosarcina pasteurii, Bacillus megaterium, Sporosarcina ureae, Proteusvulgaris, Bacillus sphaericus, Myxococcus xanthus, Proteus mirabilis,Helicobacter pylori, or variants, serotypes, mutations, or combinationsthereof.
 4. The method of claim 1, wherein the transport mediummaintains viability without promoting significant propagation of themicroorganisms.
 5. The method of claim 4, wherein the transport mediumcomprises minimal cell medium.
 6. The method of claim 1, wherein thecalcium comprises calcium chloride, calcium acetate, calcium phosphate,calcium carbonate, calcium lactate, calcium nitrate, and/or a calciumsalt.
 7. The method of claim 1, wherein the ingredients for growthcomprise one or more of water, nutrients, vitamins, minerals, aminoacids, proteins, oils, fatty acids, saccharides and polysaccharides. 8.The method of claim 1, wherein applying comprising spraying or misting.9. The method of claim 1, wherein the area is a walking path, a miningarea, tailings, gangue, a debris or waste pile, a cliff, a roadway, anaircraft landing area, a contaminated or hazardous area, or aconstruction site, or air space of a mining area, factory, manufacturingplant, or work area.
 10. The method of claim 1, wherein either or bothcompositions form a film on a surface that promotes binding of themicroorganisms.
 11. The method of claim 10, wherein the film comprises apolymer.
 12. The method of claim 1, wherein either or both compositionscontain a coloring agent.
 13. The method of claim 1, wherein either orboth compositions contain a detectable marker.
 14. The method of claim1, further comprising repeated application of either or bothcompositions to the surface.
 15. The method of claim 1, furthercomprising applying an aqueous composition to the area.
 16. A kit fordust control comprising: a first composition containing a mixture ofviable cells or spores of urease-producing microorganisms and atransport media; and a second composition containing a form of calcium,wherein either or both first and second compositions further containurea and ingredients for growth of the mixture of viable cells orspores.
 17. The kit of claim 16, wherein a mixture of viable cells orspores comprise yeast, algae, bacteria, eukaryotic cells, orrecombinantly engineered microorganisms.
 18. The kit of claim 17,wherein the bacteria comprise Sporosarcina pasteurii, Bacillusmegaterium, Sporosarcina ureae, Proteus vulgaris, Bacillus sphaericus,Myxococcus xanthus, Proteus mirabilis, Helicobacter pylori, or variants,serotypes, mutations, or combinations thereof.
 19. The kit of claim 16,wherein the transport medium maintains viability without promotingsignificant propagation of the microorganisms.
 20. The kit of claim 19,wherein the transport medium comprises minimal cell medium.
 21. The kitof claim 16, wherein the form of calcium comprises calcium chloride,calcium acetate, calcium phosphate, calcium carbonate, calcium lactate,calcium nitrate, and/or a calcium salt.
 22. The kit of claim 16, whereinthe ingredients for growth of microorganisms comprise one or more ofwater, nutrients, vitamins, minerals, amino acids, proteins, oils, fattyacids, saccharides and polysaccharides.
 23. The kit of claim 16, whereineither or both first and second compositions are aqueous.
 24. The kit ofclaim 16, wherein either or both first and second compositions furthercontain a polymer.
 25. The kit of claim 16, wherein either or both firstand second compositions further contain a coloring agent.
 26. The kit ofclaim 16, wherein either or both first and second compositions furthercontain a detectable marker.
 27. The kit of claim 16, further comprisingan apparatus for applying either or both first and second compositions.